{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:50:04Z","timestamp":1785408604682,"version":"3.56.0"},"reference-count":31,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:00:00Z","timestamp":1777593600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation Project","award":["82502717"],"award-info":[{"award-number":["82502717"]}]},{"name":"National Natural Science Foundation Project","award":["82172277"],"award-info":[{"award-number":["82172277"]}]},{"award":["82502717"],"award-info":[{"award-number":["82502717"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["82172277"],"award-info":[{"award-number":["82172277"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Beijing Natural Science Foundation Project","award":["7254305"],"award-info":[{"award-number":["7254305"]}]},{"name":"High-level Public Health Technical Talent Construction Project of Beijing Municipal Health Commission","award":["Academic Leader-02-20"],"award-info":[{"award-number":["Academic Leader-02-20"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Background: Four genotypes of human bocaviruses (HBoVs) have been identified, with only HBoV1 being detected in respiratory specimens, and with HBoV2 being the predominant human bocavirus in fecal specimens, which implies different tissue tropisms for HBoV1 and HBoV2. It is vital to determine the factors that influence the tissue tropisms. Methods: The major capsid proteins VP3 of HBoV1 and HBoV2 were expressed in eukaryotic cells. Then co-immunoprecipitation (Co-IP) and liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS) (IP-MS) was employed, along with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, to screen host proteins interacting with VP3 of different genotypes. Subsequently, in vitro pull-down assays were conducted to verify the direct virus\u2013host interaction proteins with VP3. Furthermore, molecular docking was performed to predict the interaction interfaces between viral and host proteins. Results: Through IP-MS and enrichment analyses, 50 host proteins that displayed \u226510-fold differential binding affinities between HBoV1 VP3 and HBoV2 VP3 were identified. Among these, seven were considered as high-confidence candidate interactors. Notably, SPLUNC1 and VAMP8 showed predominant expression in respiratory and intestinal tissues, respectively. Subsequent in vitro pull-down assays confirmed that SPLUNC1 specifically bound to HBoV1 VP3, whereas VAMP8 specifically interacted with HBoV2 VP3. Molecular docking analysis further revealed that the binding between SPLUNC1 with HBoV1 VP3, as well as VAMP8 with HBoV2 VP3, was stabilized by extensive hydrophobic interfaces along with specific hydrogen bonds. Conclusions: The specific interactions of host proteins SPLUNC1 with HBoV1 VP3 and VAMP8 with HBoV2 VP3, respectively, provided fundamental evidence that the distinct tissue tropisms of HBoVs may be governed by specific host factors.<\/jats:p>","DOI":"10.3390\/pathogens15050486","type":"journal-article","created":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T06:50:01Z","timestamp":1777618201000},"page":"486","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Differential Interactions of Tissue-Restricted Host Proteins SPLUNC1 and VAMP8 with VP3 of Human Bocaviruses 1 and 2"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1055-1327","authenticated-orcid":false,"given":"Ri","family":"De","sequence":"first","affiliation":[{"name":"Laboratory of Virology, Capital Center for Children\u2019s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanpeng","family":"Xu","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Capital Center for Children\u2019s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8367-8995","authenticated-orcid":false,"given":"Hanhaoyu","family":"Fu","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Capital Center for Children\u2019s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China"},{"name":"Graduate School of Peking Union Medical College, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liping","family":"Jia","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Capital Center for Children\u2019s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linqing","family":"Zhao","sequence":"additional","affiliation":[{"name":"Laboratory of Virology, Capital Center for Children\u2019s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"12891","DOI":"10.1073\/pnas.0504666102","article-title":"Cloning of a human parvovirus by molecular screening of respiratory tract samples","volume":"102","author":"Allander","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","first-page":"385","article-title":"Preliminary studies suggest that a novel parvovirus called human bocavirus is related to acute respiratory infections in pediatric patients in Beijing","volume":"5","author":"Zhao","year":"2006","journal-title":"Zhonghua Weishengwuxue Yu Mianyixue Zazhi"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1086\/595831","article-title":"A newly identified bocavirus species in human stool","volume":"199","author":"Kapoor","year":"2009","journal-title":"J. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1086\/652416","article-title":"Human bocaviruses are highly diverse, dispersed, recombination prone, and prevalent in enteric infections","volume":"201","author":"Kapoor","year":"2010","journal-title":"J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"De, R., Zhang, K.X., Wang, F., Zhou, Y.T., Sun, Y., Chen, D.M., Zhu, R.N., Guo, Q., Liu, S., and Qu, D. (2022). Human bocavirus 1 is a genuine pathogen for acute respiratory tract infection in pediatric patients determined by nucleic acid, antigen, and serology tests. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.932858"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.jcv.2010.07.016","article-title":"Human bocavirus in children: Mono-detection, high viral load and viraemia are associated with respiratory tract infection","volume":"49","author":"Christensen","year":"2010","journal-title":"J. Clin. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"104453","DOI":"10.1016\/j.jcv.2020.104453","article-title":"Human bocavirus infection and respiratory tract disease identified in a UK patient cohort","volume":"129","author":"Bagasi","year":"2020","journal-title":"J. Clin. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"T\u0103t\u0103ranu, E., Galos, F., Anchidin-Norocel, L., Axinte, R., Filip, F., Axinte, S., T\u0103t\u0103ranu, A., Terteliu, M., and Diaconescu, S. (2024). Life-threatening conditions in children with bocavirus infection-case series and mini review of the literature. Viruses, 16.","DOI":"10.3390\/v16091347"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Zhao, H., Zhao, L., Sun, Y., Qian, Y., and Dong, H. (2012). Detection of a bocavirus circular genome in fecal specimens from children with acute diarrhea in Beijing, China. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0048980"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1002\/jmv.24415","article-title":"Human bocavirus in children with acute gastroenteritis in Albania","volume":"88","author":"Iaconelli","year":"2016","journal-title":"J. Med. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Zhao, M., Zhu, R., Qian, Y., Deng, J., Wang, F., Sun, Y., Dong, H., Liu, L., Jia, L., and Zhao, L. (2016). Prevalence and phylogenetic analysis of human bocaviruses 1-4 in pediatric patients with various infectious diseases. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0160603"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"De, R., Liu, L., Qian, Y., Zhu, R., Deng, J., Wang, F., Sun, Y., Dong, H., Jia, L., and Zhao, L. (2017). Risk of acute gastroenteritis associated with human bocavirus infection in children: A systematic review and meta-analysis. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0184833"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/s00705-025-06265-x","article-title":"Exploring evidence from cells to clinics: Is human bocavirus a gastrointestinal pathogen or just a risk factor?","volume":"170","author":"Mohammadi","year":"2025","journal-title":"Arch. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1086\/592985","article-title":"CD4+ T helper cell responses against human bocavirus viral protein 2 viruslike particles in healthy adults","volume":"198","author":"Lindner","year":"2008","journal-title":"J. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4670","DOI":"10.1128\/JVI.02998-15","article-title":"Mapping Antigenic Epitopes on the Human Bocavirus Capsid","volume":"90","author":"Kailasan","year":"2016","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e02329-20","DOI":"10.1128\/JVI.02329-20","article-title":"pH-induced conformational changes of human bocavirus capsids","volume":"95","author":"Luo","year":"2021","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5880","DOI":"10.1128\/JVI.02719-09","article-title":"Human bocavirus capsid structure: Insights into the structural repertoire of the parvoviridae","volume":"84","author":"Gurda","year":"2010","journal-title":"J. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1007\/s12519-023-00697-8","article-title":"Human bocavirus 1 and 2 genotype-specific antibodies for rapid antigen testing in pediatric patients with acute respiratory infections","volume":"19","author":"De","year":"2023","journal-title":"World J. Pediatr."},{"key":"ref_19","first-page":"1289","article-title":"Human bocavirus infections are common in Beijing population indicated by sero-antibody prevalence analysis","volume":"122","author":"Zhao","year":"2009","journal-title":"Chin. Med. J. (Engl.)"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1563","DOI":"10.1080\/22221751.2019.1682949","article-title":"Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones","volume":"8","author":"Zhao","year":"2019","journal-title":"Emerg. Microbes Infect."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"eadw9158","DOI":"10.1126\/sciadv.adw9158","article-title":"Bi-handed assembly chaperones regulate protein complex assembly through an intramolecular handover mechanism","volume":"11","author":"Wu","year":"2025","journal-title":"Sci. Adv."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"774","DOI":"10.1016\/j.jmb.2007.05.022","article-title":"Inference of macromolecular assemblies from crystalline state","volume":"372","author":"Krissinel","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1042\/BST20241029","article-title":"Is the SPLUNC1-Orai1 axis a critical determinant of lung health?","volume":"53","author":"Tarran","year":"2025","journal-title":"Biochem. Soc. Trans."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1165\/rcmb.2014-0297RT","article-title":"Bactericidal\/permeability-increasing protein fold-containing family member A1 in airway host protection and respiratory disease","volume":"52","author":"Britto","year":"2015","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1002\/1522-2683(200105)22:9<1795::AID-ELPS1795>3.0.CO;2-J","article-title":"Identification of a new potential airway irritation marker, palate lung nasal epithelial clone protein, in human nasal lavage fluid with two-dimensional electrophoresis and matrix-assisted laser desorption\/ionization-time of flight","volume":"22","author":"Lindahl","year":"2001","journal-title":"Electrophoresis"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1113\/jphysiol.2011.222091","article-title":"VAMP8 is a vesicle SNARE that regulates mucin secretion in airway goblet cells","volume":"590","author":"Jones","year":"2012","journal-title":"J. Physiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e01454-19","DOI":"10.1128\/JVI.01454-19","article-title":"VAMP8 Contributes to the TRIM6-Mediated Type I Interferon Antiviral Response during West Nile Virus Infection","volume":"94","author":"Atkins","year":"2020","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4306","DOI":"10.1038\/s41467-019-11811-8","article-title":"VAMP8-mediated MUC2 mucin exocytosis from colonic goblet cells maintains innate intestinal homeostasis","volume":"10","author":"Cornick","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"426","DOI":"10.15698\/mic2017.12.605","article-title":"VAMP8 mucin exocytosis attenuates intestinal pathogenesis by Entamoeba histolytica","volume":"4","author":"Cornick","year":"2017","journal-title":"Microb. Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1089\/jir.2020.0141","article-title":"The Multifunctional Roles of Short Palate, Lung, and Nasal Epithelium Clone 1 in Regulating Airway Surface Liquid and Participating in Airway Host Defense","volume":"41","author":"Liu","year":"2021","journal-title":"J. Interferon Cytokine Res."},{"key":"ref_31","first-page":"1013","article-title":"VAMP8 is required for autophagosome-lysosome fusion","volume":"24","author":"Itakura","year":"2013","journal-title":"Mol. Biol. Cell."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/5\/486\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T06:57:10Z","timestamp":1777618630000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/5\/486"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,1]]},"references-count":31,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["pathogens15050486"],"URL":"https:\/\/doi.org\/10.3390\/pathogens15050486","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,1]]}}}